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衍生化后婴幼儿奶粉样品中左旋肉碱的测定

Determination of L-Carnitine in Infant Powdered Milk Samples after Derivatization.

作者信息

Park Jung Min, Koh Jong Ho, Kim Jin Man

机构信息

Department of Food Marketing and Safety, Konkuk University, Seoul 05029, Korea.

Department of Bio-Food Analysis, Bio-Campus, Korea Polytechnic College, Nonsan 32940, Korea.

出版信息

Food Sci Anim Resour. 2021 Jul;41(4):731-738. doi: 10.5851/kosfa.2021.e23. Epub 2021 Jul 1.

Abstract

Herein, a novel analytical method using a high-performance liquid chromatography-fluorescence detector (HPLC/FLD) is developed for rapidly measuring an L-carnitine ester derivative in infant powdered milk. In this study, solid-phase extraction cartridges filled with derivatized methanol and distilled water were used to effectively separate L-carnitine. Protein precipitation pretreatment was carried out to remove the protein and recover the analyte extract with a high recovery (97.16%-106.56%), following which carnitine in the formula was derivatized to its ester form. Precolumn derivation with 1-aminoanthracene (1AA) was carried out in a phosphate buffer using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) as the catalyst. Method validation was performed following the AOAC guidelines. The calibration curves were linear in the L-carnitine concentration range of 0.1-2.5 mg/L. The lower limit of quantitation and limit of detection of L-carnitine were 0.076 and 0.024 mg/L, respectively. The intra- and interday precision and recovery results were within the allowable limits. The results showed that our method helped reduce the sample preparation time. It also afforded higher resolution and better reproducibility than those obtained by traditional methods. Our method is suitable for detecting the quantity of L-carnitine in infant powdered milk containing a large amount of protein or starch.

摘要

本文开发了一种使用高效液相色谱 - 荧光检测器(HPLC/FLD)的新型分析方法,用于快速测定婴儿奶粉中的左旋肉碱酯衍生物。在本研究中,使用填充有衍生化甲醇和蒸馏水的固相萃取柱有效地分离左旋肉碱。进行蛋白质沉淀预处理以去除蛋白质并以高回收率(97.16% - 106.56%)回收分析物提取物,随后将配方中的肉碱衍生化为其酯形式。在磷酸盐缓冲液中使用1 - (3 - 二甲基氨基丙基) - 3 - 乙基碳二亚胺盐酸盐(EDC)作为催化剂,用1 - 氨基蒽(1AA)进行柱前衍生。按照AOAC指南进行方法验证。校准曲线在左旋肉碱浓度范围为0.1 - 2.5 mg/L时呈线性。左旋肉碱的定量下限和检测限分别为0.076和0.024 mg/L。日内和日间精密度及回收率结果均在允许范围内。结果表明,我们的方法有助于减少样品制备时间。与传统方法相比,它还具有更高的分辨率和更好的重现性。我们的方法适用于检测含有大量蛋白质或淀粉的婴儿奶粉中左旋肉碱的含量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/586b/8277179/836d13dae4f3/kosfa-41-4-731-g1.jpg

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